ATP生物发光法检测喷气燃料中真菌的污染程度  

ATP Bioluminescence Assay for Detection of Fungal Contamination in Jet Fuels

作　　者：李泽振 熊云 陈波 LI Ze-Zhen;XIONG Yun;CHEN Bo(Department of Oil,Amry Logistics University of PLA,Chongqing 401331,2.Sinopec Zhenhai Refining ＆ Chem ical Corps Room,Ningbo 315207;Chin)

机构地区：[1]中国人民解放军陆军勤务学院油料系,重庆401331 [2]驻中国石油化工股份有限公司镇海炼化分公司军代室,浙江宁波315207

出　　处：《生物技术通讯》2018年第4期525-529,共5页Letters in Biotechnology

基　　金：全军后勤科研计划重点项目(YX216J013)

摘　　要：目的:以ATP生物发光法为基础,建立一套可以快速检测喷气燃料中真菌污染程度的方法。方法:筛选喷气燃料中的枝孢霉菌、曲霉菌及青霉菌这3种主要污染真菌的ATP释放条件,比较3种ATP荧光素-萤光素酶体系的标准荧光曲线,考察ATP生物发光法与传统平板计数法计量主要污染真菌数量的相关性。结果:以表面活性剂苯扎氯铵(BAC)作为裂解液提取ATP效果最佳,最佳作用浓度与作用时间分别为0.15%和30 s,筛选得到的荧光素-萤光素酶反应体系有良好的荧光效率,其检测限可达10^(-15)mol ATP,用ATP生物发光法与传统平板计数法计量主要污染真菌的数量,两者相关系数均在0.96以上,具有良好的相关性。结论:ATP生物发光法可以用于检测喷气燃料中真菌污染程度,且能将检测时间缩短至10 min内,适用于现场快速检测,具有良好的应用前景。Objective： To establish a method based on ATP bioluminescence that can rapidly detect the degree of fungal contamination in jet fuel. Methods： The ATP release conditions of the three main contaminating fungi, Cladosporium, Amorphotheca, and Penicillium in jet fuel were screened. The standard fluorescence curves of the three ATP luciferase-luciferase systems were compared. The ATP bioluminescence method was investigated. The traditional plate count method measures the conelation of the number of major contaminating fungi. Results： Surfactant benzallkonium chloride（BAC） was chosen as the most suitablelysate of ATP extraction, and the optimal concentration and time of action were 0.15% and 30 s respectively. The screened luciferin-luciferase reaction system had good fluorescence efficiency, and its detection limit was acceptable and low as 10 15 mol ATP. The con＇elation coefficients between measurement of the number of major contaminating fungi by ATP bioluminescence method and traditional plate count methodwere all above 0.96, showing a high correlation. Conclusion： The ATP bioluminescence method can be used to detect the degree of fungal contamination in jet fuel, and can shorten the detection time to 10 min. It is suitable for on-site rapid detection and has a good application prospect.

关 键 词：ATP生物发光法 喷气燃料 真菌污染 荧光素 萤光素酶 

分 类 号：Q503[生物学—生物化学]